CAJ | 학술논문

目的利用Link-test统计方法整合分析基因芯片和蛋白质质谱分析数据寻找骨肉瘤血清蛋白标志物.方法利用基因芯片检测从骨肉瘤细胞系(MG-63、Saos-2和U-2OS)中筛选骨肉瘤候选标志基因;运用蛋白质质谱分析技术分析27个骨肉瘤患者血清标本及47个健康人的血清标本,从中筛选差异具有统计学意义的蛋白质质谱峰;采用Link-test统计方法对筛选到的骨肉瘤候选标志基因和蛋白质质谱峰进行相关性分析,得到交叉验证的骨肉瘤血清蛋白标志物;运用免疫印迹法比较一种候选血清蛋白标志物在骨肉瘤患者和健康人血清中的含量差异.结果利用基因芯片检测到653个候选骨肉瘤标志基因,蛋白质质谱分析筛选到6个蛋白质质谱峰.经Link-test统计分析,共鉴定出13个候选骨肉瘤血清蛋白标志物,其中包括细胞色素C (Cytochrome cl,CYC-1).在13个候选蛋白质标志物中,12个在骨肉瘤中表达上调,1个表达下调.免疫印迹实验发现CYC-1在骨肉瘤患者血清中的含量明显高于健康人,差异具有统计学意义.结论利用Link-test统计方法整合分析基因芯片和蛋白质质谱分析数据是寻找骨肉瘤血清蛋白标志物的有效方法,共发现13个候选骨肉瘤血清蛋白标志物.
목적 이용Link-test통계방법정합분석기인심편화단백질질보분석수거심조골육류혈청단백표지물.방법 이용기인심편검측종골육류세포계(MG-63、Saos-2화U-2OS)중사선골육류후선표지기인;운용단백질질보분석기술분석27개골육류환자혈청표본급47개건강인적혈청표본,종중사선차이구유통계학의의적단백질질보봉;채용Link-test통계방법대사선도적골육류후선표지기인화단백질질보봉진행상관성분석,득도교차험증적골육류혈청단백표지물;운용면역인적법비교일충후선혈청단백표지물재골육류환자화건강인혈청중적함량차이.결과 이용기인심편검측도653개후선골육류표지기인,단백질질보분석사선도6개단백질질보봉.경Link-test통계분석,공감정출13개후선골육류혈청단백표지물,기중포괄세포색소C (Cytochrome cl,CYC-1).재13개후선단백질표지물중,12개재골육류중표체상조,1개표체하조.면역인적실험발현CYC-1재골육류환자혈청중적함량명현고우건강인,차이구유통계학의의.결론 이용Link-test통계방법정합분석기인심편화단백질질보분석수거시심조골육류혈청단백표지물적유효방법,공발현13개후선골육류혈청단백표지물.
Objective To discover potential serum biomarkers of osteosarcoma by an integrated analysis of datasets from gene chip and surface enhanced laser desorption-ionization time-off-light mass spectrometry (SELDI-TOF-MS).Methods Gene chip was used to screen potential biomarkers of osteosarcoma from osteosarcoma cell lines (MG-63,Saos-2 and U-2OS),and SELDI-TOF-MS was used to screen protein peaks with statistically significant difference from serum samples (27 osteosarcoma cases and 47 healthy cases).The association between these two types of markers was constructed by link-test to get cross-validated serum biomarkers of osteosarcoma.Immunoblot analysis was performed to detect the expression of one candidate osteosarcoma protein maker in serums from osteosarcoma patients and members of the control group.Results Six hundred and fifty three candidate osteosarcoma gene markers were found by gene chip and 6 protein peaks were found by SELDI-TOF-MS.Thirteen differentially expressed serum markers of osteosarcoma including CYC-1 were screened by link-test analysis (12 of them were up-regulated in serum samples of the osteosarcoma patients while 1 was down-regulated ).By immunoblot,we found that CYC-1 protein was significantly up-regulated in serums from osteosarcoma patients compared to the control group.Conclusion An integrated analysis of datasets from gene chip and SELDI-TOF-MS could accelerate the discovery,screening and identification of tumor biomarkers.A total of 13 potential serum protein markers of osteosarcoma were discovered in this study.